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WO2019078381A1 - Composition pharmaceutique, composition alimentaire et additif alimentaire pour prévenir, soulager ou traiter la perte, la faiblesse et l'atrophie musculaires, contenant, à titre de principe actif, une bactérie enterococcus faecalis, le liquide de culture ou des cellules mortes de celle-ci - Google Patents

Composition pharmaceutique, composition alimentaire et additif alimentaire pour prévenir, soulager ou traiter la perte, la faiblesse et l'atrophie musculaires, contenant, à titre de principe actif, une bactérie enterococcus faecalis, le liquide de culture ou des cellules mortes de celle-ci Download PDF

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Publication number
WO2019078381A1
WO2019078381A1 PCT/KR2017/011557 KR2017011557W WO2019078381A1 WO 2019078381 A1 WO2019078381 A1 WO 2019078381A1 KR 2017011557 W KR2017011557 W KR 2017011557W WO 2019078381 A1 WO2019078381 A1 WO 2019078381A1
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Prior art keywords
enterococcus faecalis
muscular
muscle
weakness
atrophy
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PCT/KR2017/011557
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English (en)
Korean (ko)
Inventor
김택중
이명헌
이와사마사히로
한권일
김완재
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Nihon Berumu Co ltd
Korea Berm Co Ltd
University Industry Foundation UIF of Yonsei University
Original Assignee
Nihon Berumu Co ltd
Korea Berm Co Ltd
University Industry Foundation UIF of Yonsei University
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Application filed by Nihon Berumu Co ltd, Korea Berm Co Ltd, University Industry Foundation UIF of Yonsei University filed Critical Nihon Berumu Co ltd
Priority to US16/757,279 priority Critical patent/US11679134B2/en
Priority to JP2020522295A priority patent/JP7545683B2/ja
Priority to PCT/KR2017/011557 priority patent/WO2019078381A1/fr
Publication of WO2019078381A1 publication Critical patent/WO2019078381A1/fr
Anticipated expiration legal-status Critical
Priority to JP2022081930A priority patent/JP2022110113A/ja
Priority to JP2024068698A priority patent/JP2024102128A/ja
Ceased legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0087—Galenical forms not covered by A61K9/02 - A61K9/7023
    • A61K9/0095—Drinks; Beverages; Syrups; Compositions for reconstitution thereof, e.g. powders or tablets to be dispersed in a glass of water; Veterinary drenches
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/015—Inorganic compounds
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/03—Organic compounds
    • A23L29/035—Organic compounds containing oxygen as heteroatom
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/03—Organic compounds
    • A23L29/045—Organic compounds containing nitrogen as heteroatom
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/15—Vitamins
    • A23L33/155—Vitamins A or D
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
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    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A—HUMAN NECESSITIES
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    • A61K9/2004—Excipients; Inactive ingredients
    • A61K9/2013—Organic compounds, e.g. phospholipids, fats
    • A61K9/2018—Sugars, or sugar alcohols, e.g. lactose, mannitol; Derivatives thereof, e.g. polysorbates
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A61K9/20—Pills, tablets, discs, rods
    • A61K9/2004—Excipients; Inactive ingredients
    • A61K9/2022—Organic macromolecular compounds
    • A61K9/205—Polysaccharides, e.g. alginate, gums; Cyclodextrin
    • A61K9/2059—Starch, including chemically or physically modified derivatives; Amylose; Amylopectin; Dextrin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/4841—Filling excipients; Inactive ingredients
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    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
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    • A61K9/4866—Organic macromolecular compounds
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Definitions

  • the present invention relates to the use of Enterococcus
  • the present invention also relates to a novel use of the culture broth or a bacterium of the bacterium of the genus Faecalis, more specifically, to an enterococcus plant EF-2001 faecalis EF-2001), a pharmaceutical composition, a food composition and a food additive for preventing or treating muscular dysfunction, weakening and muscle atrophy, which comprises the culture liquid or a bacterium thereof as an active ingredient.
  • Diseases that cause weakness of muscle strength include sarcopenia with aging, muscle atrophy caused by imbalance of protein metabolism or muscle use, starvation, wasting disease (such as cancer), aging And acardiotrophy.
  • Sarcopenia refers to a decrease in muscle strength with a decrease in skeletal muscle mass during aging. In addition to the decrease in muscle mass, which is the most important feature of myopenia, changes in the type of muscle fiber are also observed. Type 1 and Type 2 decrease in age at the same rate, whereas Type 2 muscle fibers do not change much in the presence of myopenia, but Type 1 muscle fiber thickness decreases significantly. This myopenic disorder has been reported to cause senescence and dysfunctions among the elderly (Roubenoff R., Can. J. Appl. Physiol. 26, 78-89, 2001).
  • myopenia is caused by various factors, research on each factor is still insignificant.
  • the balance of growth hormone reduction or neurological change, changes in physical activity, changes in metabolism, sex hormone levels or fat or catabolic cytokines, and the synthesis and differentiation of proteins (Roubenoff R. and Hughes VA, J. Gerontol. A. Biol. Sci. Med. Sci. 55, M716-M724, 2000).
  • Satellite cells are small mononuclear cells located between the basement membrane and the sarcolemma of the myofiber. They are activated by stimulation, such as injury or exercise, and proliferate into myoblasts. When differentiation proceeds, they fuse with other cells to form multinuclear muscle fibers.
  • the activity of satellite cells decreases, the ability to regenerate damaged muscle or response to differentiation signals is reduced, resulting in decreased muscle formation.
  • muscle atrophy is caused by damage to muscle tissue due to lack of mechanical stimulation such as decrease of muscle use, destruction of muscles due to direct injury or physical factors, restoration of muscular cell by aging, , And disability of muscle use due to damage to the nerves that regulate muscle function (Booth FW, J Appl Physiol Respir Environ Exerc Physiol., 1982).
  • disuse atrophy occurs gradually due to loss of muscle strength due to the disability of the muscles of the affected part and the peripheral part due to disability or accident for a long time, and myasthenia due to the disease of the muscle itself gravis, dystrophy: progressive muscular dystrophy, myotonic muscular dystrophy, duchen type, Becker type, parietal lobe, facial shoulder brachial type, inflammation occurring in the muscle itself, muscular atrophy due to muscle- Amyotrophic lateral sclerosis (ALS): Lou Gehrig's disease, sphinobulbar muscular atrophy: Kennedy's disease, and the like. But also in form.
  • ALS Amyotrophic lateral sclerosis
  • the first is exercise. Exercise has been reported to increase skeletal muscle protein synthesis in the short term and to increase muscular strength and motility of the elderly. However, it is inappropriate for long-term treatment (Timothy J. Doherty, J. Appl. Physiol. 95, 1717-1727, 2003).
  • the second is the use of testosterone or anabolic steroid as a medication, but it induces menstruation in women and side effects such as prostate symptoms in men.
  • Other approved regimens include DHEA (dehydroepiandrosterone) and growth hormone, which have been reported to be therapeutically feasible at sites that include SARMs (DD Thompson, J. Musculoskelet Neuronal Interact 7, 344-345 , 2007).
  • stem cell therapy stem cell therapy which separates satellite cells and introduces them into the body after in vitro differentiation and activates satellite cells directly in the body and promotes myogenesis.
  • the best way to prevent the onset of muscular atrophy in daily life is to prevent muscle loss through continuous use of muscles, but even if muscles can not be used regardless of their will, such as injury or disability, May result in muscle atrophy caused by degeneration of the muscle. In this case, it is necessary to study countermeasures to prevent muscle atrophy by stimulating muscles or preventing muscle atrophy.
  • Reactive oxygen species are highly chemically reactive groups of molecules that are highly reactive for redox reactions that attempt to exchange electrons with nearby molecules. Due to the reactivity of these active oxygen species, surrounding substances such as protein molecules or DNA in the cell lose electrons and become oxidized to exhibit mutations, failing to perform their proper functions, and failing to function properly, Death can be induced. Oxidative stress caused by reactive oxygen species is also present in muscle cells, and oxidative stress may lead to muscle cell loss and muscle atrophy. Several stressors have been shown to affect various tissues in vivo as well as damage to muscle tissue, and have been attracting attention as various disease factors (MCKinnell IW., And Rudnicki MA., Cell, 2004).
  • Enterococcus faecalis EF-2001 Enterococcus faecalis EF-2001
  • Enterococcus faecalis EF-2001 were identified through intestinal flora screening of a 2-year-old girl.
  • the Enterococcus faecalis EF-2001 is heat-treated to kill and the cell component recovered is the Enterococcus faecalis EF-2001 microbial cell.
  • mice fed Enterococcus faecalis EF-2001 have a DSS mitigation effect in mice fed Enterococcus faecalis EF-2001 (Tadano et al., J. Japan Mibyou System association, 2011).
  • DSS Extran sulfate sodium
  • mice fed Enterococcus faecalis EF-2001 also showed a decrease in the ratio of total cholesterol and triglycerides after ingesting high-fat diets (Ku et al., Medicine and biology, 2007).
  • the Enterococcus faecalis EF-2001 was found to be a causative agent of Candida to suppress the activity of albican shows the symptoms improved and the prevention effect efficacy is started to (Ishijima et al., Med. Mycol. J, 2014), in mice by administration of antibiotics as compared to the enteric control yuikgyun is multiply rapidly and harmful bacteria is inhibited (Simohashi et al., Medicine and biology, 2002).
  • the various physiological activities of Enterococcus faecalis EF-2001 are not influenced by heat and pH due to the nature of dead cells, so they can be processed into various forms (Kan, Food industry, 2001). It is also possible to ingest a large amount of lactic acid bacteria in a small amount with a cell content of 7.5 trillion grams per gram.
  • Korean Patent No. 1,560,799 discloses a composition for treating spinal muscular atrophy comprising, as an active ingredient, hnRNP M (Heterogeneous nuclear ribonucleoprotein M) or a polynucleotide encoding the hnRNP M
  • Patent No. 1,468,123 has a disease, amyotrophic for improving or therapeutic composition comprising a mammalian umbilical cord-derived stem cells as an active ingredient is disclosed
  • Korea Patent registration No. 1385191 discloses a chicory (Cichorium intybus ) extract as an active ingredient
  • 1,349,361 discloses a pharmaceutical composition for prevention and treatment of muscle atrophy comprising Eupatorium There is chinensis . simplicifolium ) as an active ingredient.
  • the effect of the composition on the atrophy of the enterococcus faecalis, the culture solution thereof or the bacterium thereof is not known at all.
  • the present inventors have made efforts to develop a novel agent for treating muscle weakness, weakness and muscular dystrophy which is free from cytotoxicity and secured to the human body.
  • the Enterococcus faecalis EF-2001 cells are free from cytotoxicity
  • the present invention has been accomplished by discovering that it is effective to treat muscular dysfunction, weakness and muscle atrophy by inhibiting muscle cell damage and cell death caused by oxidative stress.
  • the present invention demonstrates the stability of the Enterococcus faecalis EF-2001 to the human body and inhibits muscle cell damage caused by oxidative stress, thereby remarkably inhibiting muscle weakness, weakening, and treating atrophy Respectively. Therefore, it is an object of the present invention to provide a pharmaceutical composition and a food composition for proving the efficacy of treatment of muscle wasting, weakening and muscular atrophy of Enterococcus faecalis EF-2001 dead cells and culture thereof and for preventing and treating new muscle decline, weakening and muscle atrophy And a food additive.
  • the present invention relates to the use of Enterococcus
  • the present invention also provides a pharmaceutical composition for preventing and treating muscular weakening related diseases, which comprises a culture solution thereof or a bacterium thereof as an active ingredient.
  • the present invention also provides a food composition for preventing and ameliorating muscular weakness related to muscular weakness comprising Enterococcus faecalis, a culture solution thereof or a bacterium thereof as an active ingredient.
  • the present invention also provides a food additive for preventing and ameliorating muscle weakness-related diseases of muscles containing Enterococcus faecalis, a culture solution thereof or a bacterium cell thereof as an active ingredient.
  • the present invention relates to a method of treating muscular weakness related to muscle weakness comprising administering a pharmaceutically effective amount of Enterococcus faecalis, a culture thereof or a bacterium thereof to an individual or an individual afflicted with a muscle weakness- Provide prevention methods.
  • the Enterococcus faecalis EF-2001 of the present invention faecalis EF-2001 cells do not have cytotoxicity, and exhibit remarkable effect of treating muscular dysfunction, weakening and muscle atrophy by inhibiting damage of muscle cells caused by oxidative stress. Therefore, the inventive Enterococcus faecalis,
  • the culture broth or bacterium cells thereof may be usefully used as an effective ingredient, a food composition, and a food additive for compositions for preventing or treating muscular dysfunction, weakening and muscle atrophy.
  • Enterococcus faecalis EF-2001 Entererococcus faecalis EF-2001. < / RTI >
  • FIG. 2 shows the effect of Enterococcus faecalis EF-2001 on oxidative stress-induced muscle cell damage.
  • NAC is a group treated with N-acetylcysteine, which is a positive control.
  • FIG. 3 is a graph comparing the expression of HSP70 (Heat Shock Protein) protein in muscle cells of Enterococcus faecalis EF-2001 in an oxidative stress environment.
  • HSP70 Heat Shock Protein
  • FIG. 4 shows the results of oral administration of two different concentrations of Enterococcus faecalis EF-2001 at 2 mg / kg and 30 mg / kg after surgery in which the right hindlimb nerve of the rat was excised to restrict muscle movement and induce atrophy. And an experimental plan.
  • FIG. 5 is a graph showing the results of a prophylactic treatment in which the intramuscular movement of the right hindlimb nerve of the rat was resected to induce atrophy, and then the enterococcus faecalis EF-2001 was ingested daily for 1 week before surgery
  • FIG. 3 is a view showing a muscle image of a muscle of a treatment group ingested with Enterococcus faecalis EF-2001 by using a micro-tomography apparatus after 3 weeks of operation.
  • FIG. 6 is a graph showing a comparison and analysis of the effect and the difference on the atrophy according to the ingestion of the Enterococcus faecalis EF-2001 based on the photographed image shown in FIG.
  • FIG. 7 is a graph showing the effect of Enterococcus faecalis EF-2001 cells on cell death induced by H 2 O 2 .
  • FIG. 8 is a graph comparing the expression changes of SOD1 protein in muscle cells of Enterococcus faecalis EF-2001 in an oxidative stress environment.
  • the present invention relates to the use of Enterococcus
  • the present invention also provides a pharmaceutical composition for preventing and treating muscular weakening related diseases, which comprises a culture solution thereof or a bacterium thereof as an active ingredient.
  • enterococcus microorganisms are widely available in nature and utilize carbohydrates aerobically.
  • bacteria such as enterococcus microorganisms are known to prevent damage by pathogenic microorganisms due to in vivo antagonistic action or secreted antimicrobial substances.
  • the culture of Enterococcus faecalis, its culture or its microbial cells may be used either commercially or produced by known microbial cell production methods without any toxicity and harmless to human body.
  • the culture medium refers to a culture of Enterococcus faecalis cultured in a culture medium, a culture solution, a concentrated culture solution, a culture solution, a culture filtrate, a concentrated culture filtrate, or a culture filtrate. And may be a culture filtrate obtained by removing the strain.
  • the dead cells may be prepared by heat treating the corresponding live cells or treating them with formalin or other bactericides, and the dead cells may be substantially dead.
  • the dead cells used in the present invention can be produced by the following method, but are not limited thereto:
  • Muscle weakness of the muscle means a state in which the force of one or more muscles is reduced.
  • the muscle weakness may be limited to one muscle, one side, upper or lower side of the body, or may appear throughout the body.
  • subjective muscle weakness symptoms including myopia and myalgia can be quantified in an objective way through physical examination.
  • the muscle weakness-related disease refers to all diseases that may occur due to weakness of muscle strength, such as, for example, muscular dystrophy or muscular dystrophy.
  • the myopenia refers to a gradual decrease in skeletal muscle mass due to aging, which directly leads to a decrease in muscle strength, resulting in a decrease in various body functions and a disorder.
  • the atrophy is atrophy caused by muscle atrophy caused by loss of muscle tissue caused by not using muscle, muscle atrophy caused by muscle itself, or muscle damage caused by nerve damage. Muscle atrophy caused by loss of muscle tissue caused by not using the muscles is called disuse atrophy. Muscle atrophy caused by the muscle itself is caused by myasthenia gravis or dystrophy, It is preferable that the atrophy caused by the damage of the dominant nerve is spinal muscular amyotrophy, amyotrophic lateral sclerosis, or sphinobulbar muscular atrophy.
  • the present inventors confirmed that the Enterococcus faecalis EF-2001 dead cells were not cytotoxic (see FIG. 1), and the enterococcus faecalis EF-2001 The effect of restoring the survival rate of muscle cells according to the concentration of the microbial cells was confirmed (see FIG. 2).
  • the EF-2001 cells of the Enterococcus faecalis increased the amount of HSP70 and SOD1 protein expressed in the muscle cells, (Fig. 3 and Fig.
  • the Enterococcus faecalis EF-2001 microbial cells of the present invention have no cytotoxicity and exhibit a remarkable effect of treating atrophy by inhibiting oxidative stress-induced muscle cell damage. Therefore, the Enterococcus faecalis EF -2001, a culture solution thereof, or a bacterium thereof may be usefully used as an active ingredient of a pharmaceutical composition for preventing and treating muscular weakness-related diseases such as muscle atrophy and muscle loss.
  • composition according to the present invention is administered in a pharmaceutically effective amount.
  • pharmaceutically effective amount means an amount sufficient to treat a disease at a reasonable benefit or risk ratio applicable to medical treatment, and the effective dose level will depend on the type of disease, severity, , Sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, factors including co-administered drugs, and other factors well known in the medical arts.
  • the composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered singly or in multiple doses. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without side effects, which can be easily determined by those skilled in the art.
  • the effective amount of the composition according to the present invention may vary depending on the age, sex, and body weight of the patient, and is generally in the range of 0.1 mg to 100 mg, preferably 0.2 mg to 17 mg per kg of body weight per day or every other day Or one to three times a day.
  • the dosage may be varied depending on the route of administration, the severity of obesity, sex, weight, age, etc. Therefore, the dosage is not limited to the scope of the present invention by any means.
  • composition When the composition is formulated, it is prepared using diluents or excipients such as fillers, extenders, binders, humectants, disintegrants, surfactants and the like which are usually used.
  • diluents or excipients such as fillers, extenders, binders, humectants, disintegrants, surfactants and the like which are usually used.
  • Solid preparations for oral administration include tablets, patients, powders, granules, capsules, troches and the like. These solid preparations can be prepared by adding to the lactic acid bacterium Enterococcus faecalis EF-2001 of the present invention at least one excipient, , Starch, calcium carbonate, sucrose or lactose, or gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.
  • Liquid preparations for oral administration include suspensions, solutions, emulsions or syrups. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances and preservatives may be included. have.
  • Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like.
  • non-aqueous solvent and the suspending agent examples include propylene glycol, polyethyleneglycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like.
  • injectable ester such as ethyl oleate, and the like.
  • a base for suppositories witepsol, macrogol, tween 61, cacao paper, laurin, glycerol, gelatin and the like can be used.
  • the present invention also provides a food composition for preventing and ameliorating muscular weakness related to muscular weakness comprising Enterococcus faecalis, a culture thereof or a bacterium cell thereof as an active ingredient.
  • muscle weakness related to muscle weakness refers to all diseases that may occur due to weakness of muscles, such as, for example, muscular dystrophy or muscular dystrophy.
  • the myopenia refers to a gradual decrease in skeletal muscle mass due to aging, which directly leads to a decrease in muscle strength, resulting in a decrease in various body functions and a disorder.
  • the atrophy is atrophy caused by muscle atrophy caused by loss of muscle tissue caused by not using muscle, muscle atrophy caused by muscle itself, or muscle damage caused by nerve damage. Muscle atrophy caused by loss of muscle tissue caused by not using the muscles is called disuse atrophy. Muscle atrophy caused by the muscle itself is caused by myasthenia gravis or dystrophy, It is preferable that the atrophy caused by the damage of the dominant nerve is spinal muscular amyotrophy, amyotrophic lateral sclerosis, or sphinobulbar muscular atrophy.
  • the Enterococcus faecalis EF-2001 microorganism cells or culture thereof of the present invention is free from cytotoxicity and inhibits the damage of muscle cells induced by oxidative stress, thereby exhibiting remarkable muscle reduction or prevention of muscular atrophy and therapeutic effect
  • the Enterococcus faecalis EF-2001, its culture or its dead cells of the present invention can be used as an active ingredient of a food composition for preventing and ameliorating muscular weakness related to muscle weakness.
  • the kind of the food to which the lactic acid bacterium Enterococcus faecalis EF-2001 of the present invention is added.
  • the foods to which the above substances can be added include dairy products including dairy products, meat, sausage, bread, biscuits, rice cakes, chocolate, candies, snacks, confectionery, pizza, ramen and other noodles, gums, ice cream, Beverages, alcoholic beverages and vitamin complexes, dairy products and dairy products, and includes processed foods and health functional foods in a conventional sense.
  • the food composition according to the present invention is a beverage composition
  • various flavors or natural carbohydrates such as ordinary beverages
  • natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol, and erythritol.
  • Natural flavors can be advantageously used as flavorings other than those described above.
  • the ratio of the natural carbohydrate is generally about 1 g to 20 g, preferably about 5 g to 10 g per 100 g of the composition of the present invention.
  • the present invention also provides a food additive for preventing and ameliorating muscle weakness-related diseases of muscles comprising Enterococcus faecalis, a culture solution thereof or a bacterium cell thereof as an active ingredient.
  • the food additive according to the present invention can be used as a food additive in the form of a flavoring agent such as various nutrients, vitamins, minerals (electrolyte), synthetic flavors and natural flavors, a coloring agent and a thickening agent (cheese, chocolate etc.), a pectic acid and its salt, , Organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like.
  • a flavoring agent such as various nutrients, vitamins, minerals (electrolyte), synthetic flavors and natural flavors, a coloring agent and a thickening agent (cheese, chocolate etc.), a pectic acid and its salt, , Organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like.
  • it may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks
  • the Enterococcus faecalis EF-2001 dead cells of the present invention can be added directly to food or used together with other food or food ingredients, and can be suitably used according to conventional methods.
  • the amount of the active ingredient to be mixed can be suitably determined according to the intended use (for prevention or improvement).
  • the amount of the composition of the present invention in a health functional food may be 0.1 to 90 parts by weight per total food weight.
  • the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.
  • the present invention relates to a method of treating muscular weakness related to muscle weakness comprising administering a pharmaceutically effective amount of Enterococcus faecalis, a culture thereof or a bacterium thereof to an individual or an individual afflicted with a muscle weakness- Provide prevention methods.
  • a non-human mammal such as a human or a non-human creature such as a cow, a monkey, a bird, a cat, a mouse, a rat, a hamster, a pig, a dog, a rabbit, This can be used for damaged objects.
  • the treatment can be administered orally or parenterally (for example, intramuscularly, intravenously, intraperitoneally, subcutaneously, intradermally, or topically) depending on the intended method,
  • parenterally for example, intramuscularly, intravenously, intraperitoneally, subcutaneously, intradermally, or topically
  • the severity of the disease, the form of the drug, the route of administration, and the time may be suitably selected by those skilled in the art.
  • the Enterococcus faecalis EF-2001 dead cells used in the present invention were prepared and prepared as follows.
  • the Enterococcus faecalis EF-2001 was aerobically or anaerobically cultured on a medium used for culturing a general lactic acid bacterium, and then cultured after seed culture.
  • the pH was maintained at a neutral or weakly acidic temperature of 20 to 40 ° C for 1 to 3 days.
  • the fermentation progressed so that the number of cells per g (gram) of the final raw material reached 7,500 billion won or more, it was pulverized through heat treatment and drying process.
  • Normal muscle cells (C2C12) were cultured in DMEM medium containing 5% fetal bovine serum (FBS). Normal skeletal muscle cells were cultured in a 75 cm2 plastic flask (SPL life science Co., Ltd. Korea) supplemented with 10% FBS, 7.5% NaHCO 3, 150 ⁇ g / ml, glutamine 58.4 ⁇ g / ml, and antibiotics / antimycotics ) was cultured in DMEM medium containing 4.4 ⁇ l / ml at 37 ° C and 5% CO 2. Cells were maintained in secondary culture once every 2-3 days.
  • FBS fetal bovine serum
  • Cells were removed from the bottom of the flask by treatment with 0.25% trypsin / EDTA followed by washing with CMF-PBS (calcium magnesium free-phosphate buffered saline, pH 7.2) After neutralization with the culture medium, the cells were centrifuged (1200 rpm, 5 min). The culture solution was added to the pellet of the remaining cells, and repeatedly inhaled with a sterile pipette to prepare a single cell suspension. The prepared cell suspension and trypan blue were mixed at a ratio of 1: 1 and measured using a hemocytometer on an optical microscope.
  • CMF-PBS calcium magnesium free-phosphate buffered saline, pH 7.2
  • EzCytox kit To determine the toxicity of the Enterococcus faecalis EF-2001 cells to muscle cells, an EzCytox kit was used. Specifically, normal muscle cells (C2C12) were dispensed in 96-well plates at 2 ⁇ 10 4 cells / well. The cells were cultured in an incubator at 37 ° C and 5% CO 2, and then cultured for 24 hours at various concentrations of linoleic acid at 0, 25, 50, 100, 250 and 500 ⁇ g / ml. Cell viability of normal muscle cells cultured for 24 hours was calculated using EzCytox kit to determine the cytotoxicity of Enterococcus faecalis EF-2001 cells.
  • muscle cells were placed in a 96-well plate at 2 ⁇ 10 4 cells / well and incubated for 24 hours at 37 ° C., And cultured at 5% CO 2. Then, cells of Enterococcus faecalis EF-2001 were added at concentrations of 0, 25, 50, 100, 250 and 500 ⁇ g / ml, respectively. The total volume was adjusted to 195 ⁇ l with 10% FBS- And cultured in an incubator at 37 ° C and 5% CO 2 for 24 hours. H 2 O 2 was added to the culture solution to a total volume of 200 ⁇ l at a concentration of 1 mM, and then cultured for another 120 minutes. After incubation, 10 E of EzCytox kit was added and the absorbance was measured after 1 hour to determine its effect.
  • HSP70 Heat Shock Proein
  • SOD1 protein Changes in the amount of HSP70 (Heat Shock Proein) and SOD1 protein were measured in western blot in addition group cells supplemented with Enterococcus faecalis EF-2001 cells. That is, normal muscle cells (C2C12) were dispensed in a 96-well plate at 5 ⁇ 10 4 cells / well and cultured in an incubator at 37 ° C and 5% CO 2. Thereafter, 25 ⁇ g / ml of Enterococcus faecalis EF-2001 cells were added, and the total volume was adjusted to 2 ml with 10% FBS-DMEM medium and cultured at 37 ° C. and 5% CO 2 for 24 hours.
  • C2C12 normal muscle cells
  • H 2 O 2 was added to the culture solution to a total volume of 2 ml at a concentration of 1 mM, and then cultured for another 60 minutes. Cells were disrupted to obtain proteins. HSP70 and SOD1, and beta actin antibody were used to detect expression levels of HSP70 protein and SOD1 protein using Western blot.
  • the cells of the Enterococcus faecalis EF-2001 cells can prevent the inhibition of muscle atrophy by inducing an increase in expression of HSP70 and SOD1 protein in muscle cells (FIGS. 3 and 8)
  • mice that induced muscle loss Enterococcus Lacalis EF -2001 The dead cells Comparison of ability to recover muscle loss after ingestion
  • muscle images were obtained using a micro-tomography system to measure the initial muscle volume before muscle atrophy.
  • the right hindlimb nerve of the rat was excised to restrict muscle movement and induce atrophy.
  • Enterococcus faecalis EF-2001 was orally administered to rats at two concentrations of 2 mg / kg and 30 mg / kg.
  • the prophylactic group received Enterococcus faecalis EF-2001 daily for one week prior to surgery to induce atrophy, and the treatment group received Enterococcus faecalis EF-2001 for three weeks after surgery to induce atrophy.
  • the muscle images were acquired using a micro-tomography apparatus and are shown in FIG. 5. Based on the images, the difference in the effect of the ingestion of the Enterococcus faecalis EF-2001 on muscle atrophy Respectively.
  • the present inventors confirmed that the Enterococcus faecalis EF-2001 cells were able to potently eliminate oxidative stress-induced cell death and increased radicals, thereby counteracting oxidative damage, cell death and structural changes of cells Respectively.
  • tablets were prepared by tableting according to a conventional method for producing tablets.
  • the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.
  • an injectable preparation was prepared by incorporating the aforementioned components in the amounts indicated.
  • Vitamin B6 0.5 mg
  • composition ratio of the above-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.
  • the above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The resulting solution was filtered to obtain a sterilized container, which was sealed and sterilized, And used for manufacturing.
  • compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

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Abstract

La présente invention concerne une composition pharmaceutique, une composition alimentaire et un additif alimentaire pour prévenir, soulager ou traiter la perte, la faiblesse et l'atrophie musculaires, qui contiennent toutes, à titre de principe actif, une bactérie Enterococcus faecalis, en particulier Enterococcus faecalis EF-2001, le liquide de culture ou des cellules mortes de celle-ci. Les cellules mortes d'Enterococcus faecalis EF-001 selon la présente invention manifestent des effets remarquables en termes de traitement de la perte, de la faiblesse et de l'atrophie musculaires par inhibition de l'endommagement des tissus musculaires dû au stress oxydatif, et par conséquent, les cellules mortes de la bactérie lactique Enterococcus faecalis EF-2001 ou son liquide de culture selon la présente invention peuvent être efficacement utilisés à titre de principe actif d'une composition pharmaceutique, d'une composition alimentaire et d'un additif alimentaire pour prévenir l'atrophie musculaire et la sarcopénie.
PCT/KR2017/011557 2017-10-18 2017-10-18 Composition pharmaceutique, composition alimentaire et additif alimentaire pour prévenir, soulager ou traiter la perte, la faiblesse et l'atrophie musculaires, contenant, à titre de principe actif, une bactérie enterococcus faecalis, le liquide de culture ou des cellules mortes de celle-ci Ceased WO2019078381A1 (fr)

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US16/757,279 US11679134B2 (en) 2017-10-18 2017-10-18 Pharmaceutical composition, food composition and food additive for preventing, alleviating or treating muscle loss, weakness and atrophy, containing, as active ingredient, Enterococcus faecalis, culture liquid thereof or dead cells thereof
JP2020522295A JP7545683B2 (ja) 2017-10-18 2017-10-18 エンテロコッカス・フェカーリス、その培養液またはその死菌体を有効成分として含有する筋肉の減退、低下及び筋萎縮の予防、改善または治療用の薬学組成物、食品組成物及び食品添加剤
PCT/KR2017/011557 WO2019078381A1 (fr) 2017-10-18 2017-10-18 Composition pharmaceutique, composition alimentaire et additif alimentaire pour prévenir, soulager ou traiter la perte, la faiblesse et l'atrophie musculaires, contenant, à titre de principe actif, une bactérie enterococcus faecalis, le liquide de culture ou des cellules mortes de celle-ci
JP2022081930A JP2022110113A (ja) 2017-10-18 2022-05-18 エンテロコッカス・フェカーリス、その培養液またはその死菌体を有効成分として含有する筋肉の減退、低下及び筋萎縮の予防、改善または治療用の薬学組成物、食品組成物及び食品添加剤
JP2024068698A JP2024102128A (ja) 2017-10-18 2024-04-19 エンテロコッカス・フェカーリス、その培養液またはその死菌体を有効成分として含有する筋肉の減退、低下及び筋萎縮の予防、改善または治療用の薬学組成物、食品組成物及び食品添加剤

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JP7545683B2 (ja) * 2017-10-18 2024-09-05 コリア ベルム シーオー．，エルティーディー． エンテロコッカス・フェカーリス、その培養液またはその死菌体を有効成分として含有する筋肉の減退、低下及び筋萎縮の予防、改善または治療用の薬学組成物、食品組成物及び食品添加剤

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